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生物样品中几丁质酶活性的测定。

Measurement of Chitinase Activity in Biological Samples.

作者信息

Amick Alyssa K, Liu Qing, Gautam Samir, Chupp Geoffrey, Dela Cruz Charles S, Sharma Lokesh

机构信息

Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine.

Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine;

出版信息

J Vis Exp. 2019 Aug 22(150). doi: 10.3791/60159.

Abstract

Chitinases are the enzymes that cleave chitin. Even in the absence of chitin, mammalians have significant amounts of chitinases present in the body including at baseline. The precise role of chitinase is not known, however it was believed to play important role in digestion and host defense against chitin-containing food and pathogens, respectively. Recent work, including ours, has shown an important role of chitinase and chitinase-like proteins in host immunity and allergic diseases. Importantly, chitinase activities serve as important biomarkers of disease severity in a wide-range of diseases including type 2 inflammatory diseases such as asthma and pulmonary fibrosis. Similarly, patients with genetic disorders like Gaucher disease have significantly elevated chitinase levels, which not only correlate with disease severity but also serve as a reliable biomarker for therapeutic effectiveness. The protocol outlined here describes a simple, quick, and straightforward way to measure chitinase activity in BAL or serum samples of mice and can be widely adapted to human subjects and other model organisms due to the highly conserved nature of the enzymes.

摘要

几丁质酶是能够裂解几丁质的酶。即使在没有几丁质的情况下,哺乳动物体内也存在大量几丁质酶,包括在基线水平时。几丁质酶的确切作用尚不清楚,不过据信它分别在消化以及宿主抵御含几丁质的食物和病原体方面发挥重要作用。包括我们的研究在内,近期的研究工作表明几丁质酶和几丁质酶样蛋白在宿主免疫和过敏性疾病中发挥重要作用。重要的是,几丁质酶活性在包括哮喘和肺纤维化等2型炎症性疾病在内的多种疾病中是疾病严重程度的重要生物标志物。同样,患有戈谢病等遗传疾病的患者几丁质酶水平显著升高,这不仅与疾病严重程度相关,还可作为治疗效果的可靠生物标志物。此处概述的方案描述了一种简单、快速且直接的方法来测量小鼠支气管肺泡灌洗(BAL)液或血清样本中的几丁质酶活性,并且由于这些酶具有高度保守的性质,该方法可广泛应用于人类受试者和其他模式生物。

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